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目的分离纯化黄鳝血清免疫球蛋白,制备其兔抗血清,并检测抗血清的特异性。方法用Protein A亲和层析的方法纯化黄鳝血清免疫球蛋白,通过SDS-聚丙烯酰胺凝胶电泳检测蛋白的纯度,免疫大耳白兔制备抗血清,利用免疫双扩散检测抗血清的效价,通过western blotting检测抗血清的特异性。结果纯化了黄鳝血清免疫球蛋白,免疫双扩散法测定兔抗黄鳝免疫球蛋白血清效价为1∶32,western blotting结果显示抗血清具有很好的特异性。结论成功纯化了黄鳝免疫球蛋白,制备了兔抗黄鳝IgM抗血清,为建立黄鳝的血清学检测系统奠定了基础。
Objective To isolate and purify eel serum immunoglobulin, prepare its rabbit antiserum and detect its specificity. Methods The serum immunoglobulin of the eel was purified by Protein A affinity chromatography. The purity of the protein was detected by SDS-polyacrylamide gel electrophoresis. Immunized rabbits were used to prepare antiserum. The titer of antiserum was detected by double immunodiffusion The specificity of the antiserum was tested by western blotting. Results The serum immunoglobulin of the eel was purified. The titer of immunoglobulin of rabbit eel against Monopterus albus was 1:32 by immunodouble diffusion method. The result of western blotting showed that antiserum had good specificity. Conclusion The immunoglobulin of Monopterus albus was successfully purified and the antiserum of rabbit anti-eel IgM was prepared, which laid a foundation of establishing the serological detection system of Monopterus albus.